Comparative transient expression assay analysis of hycu-hr6- and IE1-dependent regulation of baculovirus gp64 early promoters in three insect cell lines

Nagai, Setsuko; Alves, Cristiano A. Felipe; Kobayashi, Michihiro; Ikeda, Motoko

doi:10.1016/j.virusres.2010.08.025

Cited by 4 publications

(2 citation statements)

References 37 publications

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“…Another study showed that ires mediates target gene expression in HeLa and CHO cells, with 20-50% of expression due to the promoter, and this phenomenon also occurs in insect Sf9 cells [26]. The promoter p64, which mediates GP64 expression, is a typical very early promoter (12-24 h post infection); therefore, this result also used the early promoter pie1 to reduce competition and improve display efficiency [27]. In addition, the pie1 promoter exhibits high expression activity in both insect and mammalian cells, which could facilitate gene expression by recombinant baculovirus vectors in non-host cells [28][29][30].…”

Section: Discussionmentioning

confidence: 99%

Impact of Molecular Modification on the Efficiency of Recombinant Baculovirus Vector Invasion to Mammalian Cells and Its Immunogenicity in Mice

Zheng

Pan

Wang

et al. 2022

Viruses

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The baculovirus display system (BDS), an excellent eukaryotic surface display technology that offers the advantages of safety, efficiency, and economy, is widely used in biomedicine. A previous study using rBacmid-Δgp64-ires-gp64 expressed in low copy numbers of the gp64 gene achieved high-efficiency expression and co-display of three fluorescent proteins (GFP, YFP, and mCherry). However, low expression of GP64 in recombinant baculoviruses also reduces the efficiency of recombinant baculovirus transduction into mammalian cells. In addition, the baculovirus promoter has no expression activity in mammalian cells and thus cannot meet the application requirements of baculoviral vectors for the BDS. Based on previous research, this study first determined the expression activity of promoters in insect Spodoptera frugiperda 9 cells and mammalian cells and successfully screened the very early promoter pie1 to mediate the co-expression of multiple genes. Second, utilizing the envelope display effect of the INVASIN and VSVG proteins, the efficiency of transduction of recombinant baculovirus particles into non-host cells was significantly improved. Finally, based on the above improvement, a recombinant baculovirus vector displaying four antigen proteins with high efficiency was constructed. Compared with traditional BDSs, the rBacmid-Δgp64 system exhibited increased display efficiency of the target protein by approximately 3-fold and induced an approximately 4-fold increase in the titer of serum antibodies to target antigens in Bal B/c mice. This study systematically explored the application of a new multi-gene co-display technology applicable to multi-vaccine research, and the results provide a foundation for the development of novel BDS technologies.

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Section: Discussionmentioning

confidence: 99%

Impact of Molecular Modification on the Efficiency of Recombinant Baculovirus Vector Invasion to Mammalian Cells and Its Immunogenicity in Mice

Zheng

Pan

Wang

et al. 2022

Viruses

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“…GP64 is required for binding of the BV to a host cell and subsequent fusion of the BV envelope and endosomal membrane during the penetration phase of the virus infection cycle (Volkman et al, 1984). During a later phase of baculovirus infection, the amount of GP64 that is found in the host cell membrane is related to the efficiency of BV production and rate of systemic infection in insects (Nagai et al, 2011).…”

Section: Industrial Entomologymentioning

confidence: 99%

Effect of 1-deoxynojirimycin on the Replication of Baculoviruses, Bombyx Mori Nucleopolyhedrovirus and Autographa Californica Multiple Nucleopolyhedrovirus

Kang¹,

Park²,

Cho³

et al. 2011

International Journal of Industrial Entomology

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1-Deoxynojirimycin (DNJ) is an alkaloid that is found at relatively high concentrations in mulberry leaf and tissues of the silkworm, Bombyx mori. DNJ is a well known inhibitor of α-glucosidase, an enzyme that is involved in the early stages of the N-linked glycoprotein synthesis pathway. α-Glucosidase activity in the cell extract from B. mori-derived Bm5 cells showed approximately 40-fold less sensitivity to DNJ than αglucosidase activity in the cell extract from Spodoptera frugiperda-derived Sf9 cells. The replication of B. mori nucleopolyhedrovirus (BmNPV) was not inhibited when it was propagated in BmN cells that were grown in medium containing up to 10 mM DNJ. In contrast, the replication of Autographa californica multiple NPV (AcMNPV) was reduced by 67% when it was propagated in Sf9 cells that were grown in medium containing 10 mM DNJ. The viability of Bm5 and Sf9 cells that were grown in medium containing up to 10 mM DNJ was not affected. Our results suggested that the reduced replication of AcMNPV was the result of the higher sensitivity of α-glucosidase activity in Sf9 cells to DNJ.

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Antiviral activity and specific modes of action of bacterial prodigiosin against Bombyx mori nucleopolyhedrovirus in vitro

Zhou

Zeng

Liu

et al. 2015

Appl Microbiol Biotechnol

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Prodigiosin, the tripyrrole red pigment, is a bacterial secondary metabolite with multiple bioactivities; however, the antiviral activity has not been reported yet. In the present study, we found the antiviral activity of bacterial prodigiosin on Bombyx mori nucleopolyhedrovirus (BmNPV)-infected cells in vitro, with specific modes of action. Prodigiosin at nontoxic concentrations selectively killed virus-infected cells, inhibited viral gene transcription, especially viral early gene ie-1, and prevented virus-mediated membrane fusion. Under prodigiosin treatment, both progeny virus production and viral DNA replication were significantly inhibited. Fluorescent assays showed that prodigiosin predominantly located in cytoplasm which suggested it might interact with cytoplasm factors to inhibit virus replication. In conclusion, the present study clearly indicates that prodigiosin possesses significant antiviral activity against BmNPV.

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Comparative transient expression assay analysis of hycu-hr6- and IE1-dependent regulation of baculovirus gp64 early promoters in three insect cell lines

Cited by 4 publications

References 37 publications

Impact of Molecular Modification on the Efficiency of Recombinant Baculovirus Vector Invasion to Mammalian Cells and Its Immunogenicity in Mice

Impact of Molecular Modification on the Efficiency of Recombinant Baculovirus Vector Invasion to Mammalian Cells and Its Immunogenicity in Mice

Effect of 1-deoxynojirimycin on the Replication of Baculoviruses, Bombyx Mori Nucleopolyhedrovirus and Autographa Californica Multiple Nucleopolyhedrovirus

Antiviral activity and specific modes of action of bacterial prodigiosin against Bombyx mori nucleopolyhedrovirus in vitro

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